BMPO (Spin trapping reagent)自旋捕获剂

BMPO;DEPMPO;EMPO;Spin trapping reagent自旋捕获剂;电子顺磁共振波谱(EPR);CAS: 387334-31-8;

产品信息

产品名称

产品编号 CAS NO. 规格

BMPO (Spin trapping reagent)自旋捕获剂

MX4722-5MG 387334-31-8 5mg
BMPO (Spin trapping reagent)自旋捕获剂 MX4722-10MG 387334-31-8 10mg

BMPO (Spin trapping reagent)自旋捕获剂 MX4722-50MG 387334-31-8 50mg

产品描述

BMPO(5-tert-butoxycarbonyl 5-methyl-1-pyrroline N-oxide)是一种新型高效且高稳定型的硝酮自旋捕获剂(Nitrone Spin Trapping Reagent),由美国威斯康星医学院Kalyanaraman教授的实验室团队开发,非常适合用于特异性检测和鉴定体内外形成的硫自由基、羟基自由基(•OH)和超氧阴离子自由基(•O2-),通过形成用电子顺磁共振波谱(EPR)可区分的加合物来测定[1-2]。

BMPO具有以下特点:

①特异性高,半衰期长:其它的硝酮自旋捕获剂比如:DMPO,难以轻易区分超氧阴离子和羟基自由基,因产生的DMPO-超氧加合物(半衰期t½=45s)瞬间衰减产生DMPO-羟基加合物。与最近开发的自旋捕获剂:DEPMPO和EMPO类似,BMPO-超氧加合物不会衰减生成羟基加合物,但是,BMPO-超氧加合物的半衰期最长(t½=23min)。

②产品稳定性高:DEPMPO和EMPO是液体自旋捕获剂,通常被硝基氧杂质污染,且效期有限。BMPO是固体的环形硝酮,通过结晶以高度纯化的状态提供,能够保存更长的周期,且不用担心降解。

③更高的信噪比:BMPO衍生的加合物在各自的EPR光谱中具有更高的信噪比,使其更适合用于检测细胞悬浮液中的亚硫酸、羟基和甲基自由基。

④高水溶性:水溶性好,更利于水相体系自由基的研究,尤其是生物体系的自由基研究。

产品特性

1) CAS NO:387334-31-8

2) 化学名:3,4-dihydro-2-methyl-1,1-dimethylethyl ester-2H-pyrrole-2-carboxylic acid-1-oxide

3) 同义名:5-tert-butoxycarbonyl 5-methyl-1-pyrroline N-oxide; BocMPO;

4) 分子式:C10H17NO3

5) 分子量:199.2

6) 纯度:≥98%

7) 外观:结晶或结晶性粉末

8) 溶解性:溶于水、PBS(pH 7.2, 10mg/ml)、DMSO(25mg/ml)、无水乙醇(25mg/ml)

8)化学结构式:

保存与运输方法

保存:-20ºC干燥保存,2年有效。 

运输:冰袋运输。

注意事项

为了您的安全和健康,请穿实验服并戴一次性手套操作。

参考文献

1. Zhao, H., Joseph, J., Zhang, H., et al. Synthesis and biochemical applications of a solid cyclic nitrone spin trap: A relatively superior trap for detecting superoxide anions and glutathiyl radicals. Free Radic. Biol. Med. 31(5), 599-606 (2001).

2. Khan, N., Wilmont, C.M., Rosen, G.M., et al. Spin traps: In vitro toxicity and stability of radical adducts. Free Radical Biology & Medicine 34(11), 1473-1481 (2003).

应用示例(仅作参考)

①黄嘌呤-黄嘌呤氧化酶体系内超氧阴离子(O2⋅−)生成的测定:Typically, xanthine oxidase (0.04 U/mL) was added to pH ∼7.4 sodium phosphate buffer (50 mM) containing DTPA (1 mM) and hypoxanthine (0.5–400 μM, final concentration) to achieve rates of O2⋅− formation that ranged from 0.1 to 6.0 μM/min. We estimated the superoxide formation rate by monitoring the SOD-inhibitable reduction of ferricytochrome c (80 μM) at room temperature. Spin trapping was performed by addition of 100 mM BMPO in pH ∼7.4 phosphate-buffered saline (PBS; 50 mM) containing 1 mM DTPA to the solution of hypoxanthine and xanthine oxidase to achieve a final BMPO concentration of 50 mM in the reaction mixture. EPR spectra were recorded 10 min after mixing reagents. The half-life of BMPO-OOH at ambient temperature is reported to be ∼23 min. Solutions for control experiments contained SOD (30 U/mL).

Figure 2 Comparison of CW and rapid-scan spectra of BMPO-OOH in solution with a O2⋅− production rate of 0.1μM/min, recorded 10 min after mixing reagents. The O2⋅− was produced by a hypoxanthine/xanthine oxidase mixture. The concentration of BMPO-OOH is ∼0.3μM. (A) CW spectrum obtained with 55 G sweep width, 0.75 G modulation amplitude, single 30 s scan, 15 ms conversion time, 10 ms time constant, and 20 mW (B1 = 170 mG) microwave power. (B) Deconvolved rapid-scan spectrum obtained with 55 G scan width, 51 kHz scan frequency, and 53 mW (B1 = 250 mG) microwave power. Segments consisting of 12 sinusoidal cycles were averaged 100 k times, with a total data acquisition time of ∼30 s.

2.文献来源:Wang, Z., Zhang, Y., Ju, E.et al. Biomimetic nanoflowers by self-assembly of nanozymes to induce intracellular oxidative damage against hypoxic tumors.Nat Commun 9,3334 (2018). https://doi.org/10.1038/s41467-018-05798-x 

①缺氧下的ESR测定(O2•−,•OH):For O2•− detection, the pre-deoxidized PBS (pH 5.0, 25 mM) contained 25 mM BMPO, 20 μg mL−1 MnO2@PtCo nanoflowers, 100 μM H2O2, 50% DMSO was prepared. After incubation of 5 min, ESR spectra were recorded. For •OH detection, the pre-deoxidized PBS (pH 5.0, 25 mM) contained 25 mM BMPO, 20 μg mL−1 MnO2@PtCo nanoflowers, 100 μM H2O2, 0.25 U mL−1 SOD was prepared. After incubation of 5 min, ESR spectra were recorded. The following instrument settings were used for collecting ESR spectra: 1 G field modulation, 100 G scan range, and 20 mW microwave power.

Fig f ESR spectra of BMPO/•OOH adducts from different groups in the hypoxic H2O2 (100 μM) condition upon addition of DMSO.g ESR spectra of BMPO/•OH adducts were collected from different samples in the hypoxic H2O2 (100 μM) condition upon addition of SOD. Data were presented as mean ± s.d. (n= 5).

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